GENBIO

CHAPTER 1: Introduction to Cells

Essential Concepts
 Cells are the fundamental units of life. All present-day cells are believed to have evolved from an ancestral cell
that existed more than 3 billion years ago.
 All cells, and hence all living things, grow, convert energy from one form to another, sense and respond to their
environment, and reproduce themselves.
 All cells are enclosed by a plasma membrane that separates the inside of the cell from the environment. All cells
contain DNA as a store of genetic information and use it to guide the synthesis of RNA molecules and of
proteins.
 Cells in a multicellular organism, though they all contain the same DNA, can be very different. They turn on
different sets of genes according to their developmental history and to cues they receive from their
environment.
 Cells of animal and plant tissues are typically 5–20 mm in diameter and can be seen with a light microscope,
which also reveals some of their internal components, or organelles.
 The electron microscope permits the smaller organelles and even individual large molecules to be seen, but
specimens require elaborate preparation and cannot be viewed alive.
 The simplest of present-day living cells are procaryotes: although they contain DNA, they lack a nucleus and
other organelles and probably resemble most closely the ancestral cell.
 Different species of procaryotes are diverse in their chemical capabilities and inhabit an amazingly wide range of
habitats. Two fundamental evolutionary subdivisions are recognized: bacteria and archaea.
 Eucaryotic cells possess a nucleus and other organelles not found in procaryotes. They probably evolved in a
series of stages. An important step appears to have been the acquisition of mitochondria, which are thought to
have originated from bacteria engulfed by an ancestral eucaryotic cell.
 The nucleus is the most prominent organelle in most plant and animal cells. It contains the genetic information
of the organism, stored in DNA molecules. The rest of the cell’s contents, apart from the nucleus, constitute the
cytoplasm.
 The cytoplasm includes all of the cell’s contents outside the nucleus. It contains a variety of membrane-enclosed
organelles with specialized chemical functions. Mitochondria carry out the oxidation of food molecules. In plant
cells, chloroplasts perform photosynthesis. The endoplasmic reticulum, the Golgi apparatus, and lysosomes
permit cells to synthesize complex molecules for export from the cell and for insertion in cell membranes, and to
import and digest large molecules.
 Outside the membrane-enclosed organelles in the cytoplasm is the cytosol, a concentrated mixture of large and
small molecules that carry out many essential biochemical processes.
 The cytoskeleton extends throughout the cytoplasm. This system of protein filaments is responsible for cell
shape and movement and for the transport of organelles and molecules from one location to another in the
cytoplasm.
 Free-living, single-celled eucaryotic microorganisms include some of the most complex eucaryotic cells known,
and they are able to swim, mate, hunt, and devour food.
 An animal, plant, or fungus consists of diverse eucaryotic cell types all derived from a single fertilized egg cell;
the number of such cells cooperating to form a large multicellular organism such as a human runs into
thousands of billions.
 Biologists have chosen a small number of model organisms to study closely. These include the bacterium E. coli,
brewer’s yeast, a nematode worm, a fly, a small plant, a fish, a mouse, and the human species itself.
 Although the minimum number of genes needed for a viable cell is less than 400, most cells contain significantly
more. Yet even such a complex organism as a human has only about 24,000 protein-coding genes—twice as
many as a fly and seven times as many as E. coli.

CHAPTER 2: Chemical Components of Cells

EssenTial Concepts
  Living cells obey the same chemical and physical laws as nonliving things. Like all other forms of matter, they are
composed of atoms, which are the smallest units of a chemical element that retains the distinctive chemical
properties of that element.
 Atoms are made up of smaller particles. The nucleus of an atom contains protons, which are positively charged,
and uncharged neutrons. The nucleus is surrounded by a cloud of negatively charged electrons.
 The number of electrons in an atom is equal to the number of protons in its nucleus. The nuclei of different
isotopes of the same element contain the same number of protons but different numbers of neutrons.
 Living cells are made up of a limited number of elements, four of which—C, H, N, O—make up 96.5% of their
mass.
 The chemical properties of an atom are determined by the number and arrangement of its electrons. An atom is
most stable when all of its electrons are at their lowest possible energy level and when each electron shell is
completely filled.
 Chemical bonds form between atoms as electrons move to reach a more stable arrangement. Clusters of two or
more atoms held together by chemical bonds are known as molecules.
 When an electron jumps from one atom to another, two ions of opposite charge are generated; ionic bonds can
then arise by the mutual attraction of these charged atoms.
 A covalent bond consists of a pair of electrons shared between two adjacent atoms. If two pairs of electrons are
shared, a double bond is formed.
 Living organisms contain a distinctive and restricted set of small carbon-based molecules that are essentially the
same for every living species. The main categories are sugars, fatty acids, amino acids, and nucleotides.
 Sugars are a primary source of chemical energy for cells and can be incorporated into polysaccharides for energy
storage.
 Fatty acids are also important for energy storage, but their most essential function is in the formation of cell
membranes.
 The vast majority of the dry mass of a cell consists of macromolecules, formed as polymers of sugars, amino
acids, or nucleotides.
 Macromolecules are intermediate in both size and complexity between small molecules and cell organelles.
They have many remarkable properties that are not easily deduced from the subunits from which they are
made.
 The remarkably diverse and versatile class of macromolecules known as proteins are polymers formed from
amino acids.
 Nucleotides play a central part in energy transfer and are the subunits from which the informational
macromolecules, RNA and DNA, are made.
 Protein, RNA, and DNA molecules are synthesized from subunits by repetitive condensation reactions. Each of
these biological macromolecules has a unique sequence of subunits.
 Weak noncovalent bonds form between different regions of a macromolecule. These can cause the
macromolecule to fold into a unique three-dimensional shape (conformation) with a special chemistry, as seen
most conspicuously in proteins.

CHAPTER 3: Energy, Catalysis and Biosynthesis

essenTial ConCePTs
 Living organisms are able to exist because of a continual input of energy. Part of this energy is used to carry out
essential functions—reactions that support cellular metabolism, growth, and reproduction—and the remainder
is lost in the form of heat.
 The primary source of energy for most living organisms is the sun. Plants, algae, and photosynthetic bacteria use
solar energy to produce organic molecules from carbon dioxide. Animals obtain food by eating plants or by
eating animals that feed on plants.
 Each of the many hundreds of chemical reactions that occur in a cell is specifically catalyzed by an enzyme. Large
numbers of different enzymes work in sequence to form chains of reactions, called metabolic pathways, each
performing a different function in the cell.
  Catabolic reactions break down food molecules through oxidative pathways and release energy. Anabolic
reactions generate the many complex molecules needed by the cell, and they require an energy input. In animal
cells, both the building blocks and the energy required for the anabolic reactions are obtained by catabolism.
 Enzymes catalyze reactions by binding to particular substrate molecules in a way that lowers the activation
energy required for making and breaking specific covalent bonds.
 The rate at which an enzyme catalyzes a reaction depends on how rapidly it finds its substrates and how quickly
the product forms and then diffuses away. These rates vary widely from one enzyme to another, and they can
be measured after mixing purified enzymes and substrates together under a set of defined conditions.
 The only chemical reactions possible are those that increase the total amount of disorder in the universe. The
free-energy change for a reaction, DG, measures this disorder, and it must be less than zero for a reaction to
proceed spontaneously.
 The free-energy change for a chemical reaction, DG, depends on the concentrations of the reacting molecules,
and it may be calculated from these concentrations if the equilibrium constant (K) of the reaction (or the
standard free-energy change, DG°, for the reactants) is known.
 Equilibrium constants govern all of the associations (and dissociations) that occur between macromolecules and
small molecules in the cell. The larger the binding energy between two molecules, the larger the equilibrium
constant and the more likely that these molecules will be found bound to each other.
 By creating a reaction pathway that couples an energetically favorable reaction to an energetically unfavorable
one, enzymes can make otherwise impossible chemical transformations occur.
 A small set of activated carrier molecules, particularly ATP, NADH, and NADPH, plays a central part in these
coupling events. ATP carries high-energy phosphate groups, whereas NADH and NADPH carry high-energy
electrons. • Food molecules provide the carbon skeletons for the formation of larger molecules. The
covalent bonds of these larger molecules are typically produced in reactions that are coupled to energetically
favorable bond changes in activated carrier molecules such as ATP and NADPH.

CHAPTER 4: Protein Structure and Function

eSSenTial concepTS
 Living cells contain an enormously diverse set of protein molecules, each made as a linear chain of amino acids
covalently linked together.
 Each type of protein has a unique amino acid sequence that determines both its three-dimensional shape and its
biological activity.
 The folded structure of a protein is stabilized by noncovalent interactions between different parts of the
polypeptide chain.
 Hydrogen bonds between neighboring regions of the polypeptide backbone often give rise to regular folding
patterns, known as a helices and b sheets.
 The structure of many proteins can be subdivided into smaller globular regions of compact three-dimensional
structure, known as protein domains.
 The biological function of a protein depends on the detailed chemical properties of its surface and how it binds
to other molecules, called ligands.
 When a protein catalyzes the formation or breakage of covalent bonds in a ligand, the protein is called an
enzyme and the ligand is called a substrate.
 At the active site of an enzyme, the amino acid side chains of the folded protein are precisely positioned so that
they favor the formation of the high-energy transition states that the substrates must pass through to be
converted to product.
 The three-dimensional structure of many proteins has evolved so that the binding of a small ligand can induce a
significant change in protein shape.
 Most enzymes are allosteric proteins that can exist in two conformations that differ in catalytic activity, and the
enzyme can be turned on or off by ligands that bind to a distinct regulatory site to stabilize either the active or
the inactive conformation.
 The activities of most enzymes within the cell are strictly regulated. One of the most common forms of
regulation is feedback inhibition, in which an enzyme early in a metabolic pathway is inhibited by its binding to
one of the pathway’s end products.
  Many thousands of proteins in a typical eucaryotic cell are regulated either by cycles of phosphorylation and
dephosphorylation, or by the binding and hydrolysis of GTP by a GTP-binding protein.
 The hydrolysis of ATP to ADP by motor proteins produces directed movements in the cell.
 Highly efficient protein machines are formed by assemblies of allosteric proteins in which conformational
changes are coordinated to perform complex cellular functions.
 A regulatory protein code based on the covalent modification of multiple amino acid side chains allows each cell
to control the location and assembly of its protein complexes.
 Starting from crude cell homogenates, individual proteins can be obtained in pure form by using a series of
chromatography steps. Purification allows the detailed properties of a protein to be revealed by biochemical
techniques and its exact three-dimensional structure to be determined.

CHAPTER 5: DNA and Chromosomes

eSSenTial concePTS
 Life depends on the stable and compact storage of genetic information.
 Genetic information is carried by very long DNA molecules and encoded in the linear sequence of nucleotides A,
T, G, and C.
 Each molecule of DNA is a double helix composed of a pair of complementary strands of nucleotides held
together by hydrogen bonds between G-C and A-T base pairs.
 A strand of DNA has a chemical polarity due to the linkage of alternating sugars and phosphates in its backbone.
The two strands of the DNA double helix run antiparallel—that is, in opposite orientations.
 The genetic material of a eucaryotic cell is contained in a set of chromosomes, each formed from a single,
enormously long DNA molecule that contains many genes.
 When a protein-coding gene is expressed, part of its nucleotide sequence is copied into RNA, which then directs
the synthesis of a specific protein.
 The DNA that forms each eucaryotic chromosome contains, in addition to genes, many replication origins, one
centromere, and two telomeres. These sequences ensure that the chromosome can be replicated efficiently and
passed on to daughter cells
 Chromosomes in eucaryotic cells consist of DNA tightly bound to a mass of specialized proteins. These proteins
fold the DNA into a compact form. The complex of DNA and protein in chromosomes is called chromatin.
 The most abundant chromosomal proteins are the histones, which pack DNA into a repeating array of DNA–
protein particles called nucleosomes.
 Nucleosomes pack together, with the aid of histone H1 molecules, to form a 30-nm fiber. This fiber is generally
coiled and folded, producing more compact chromatin structures.
 Chromatin structure is dynamic: by temporarily decondensing its structure—using chromatin remodeling
complexes and enzymes that covalently modify histone tails—the cell can ensure that proteins involved in gene
expression, replication, and repair have rapid, localized access to the necessary DNA sequences.
 Some forms of chromatin have a pattern of histone tail modification that causes the DNA to become so highly
compacted that the packaged genes cannot be expressed to produce RNA and protein.
 Chromatin structure can be transmitted from one cell generation to the next, producing a form of epigenetic
inheritance that helps a cell to remember the state of gene expression in its parent cell.

CHAPTER 6: DNA Replication, Repair, and Recombination

ESSEnTIaL COnCEPTS
 The ability of a cell to maintain order in a chaotic environment depends on the accurate duplication of the vast
quantity of genetic information carried in its DNA.
 Each of the two DNA strands can act as a template for the synthesis of the other strand. A DNA double helix thus
carries the same information in each of its strands.
 A DNA molecule is duplicated (replicated) by the polymerization of new complementary strands using each of
the old strands of the DNA double helix as a template. Two identical DNA molecules are formed, enabling the
genetic information to be copied and passed on from cell to daughter cell and from parent to offspring.
  As a DNA molecule replicates, its two strands are pulled apart to form one or more Y-shaped replication forks.
DNA polymerase enzymes, situated at the fork, produce a new complementary DNA strand on each parental
strand.
 DNA polymerase replicates a DNA template with remarkable fidelity, making less than one error in every 107
bases read. This accuracy is made possible, in part, by a proofreading process in which the enzyme removes its
own polymerization errors as it moves along the DNA.
 Because DNA polymerase synthesizes new DNA in only one direction, only the leading strand at the replication
fork can be synthesized in a continuous fashion. On the lagging strand, DNA is synthesized in a discontinuous
backstitching process, producing short fragments of DNA that are later joined by the enzyme DNA ligase to
complete that DNA strand.
 DNA polymerase is incapable of starting a new DNA chain from scratch. Instead, DNA synthesis is primed by an
RNA polymerase called primase, which makes short lengths of RNA (primers) that are elongated by DNA
polymerase. The primers are subsequently erased and replaced with DNA.
 DNA replication requires the cooperation of many proteins; these form a multienzyme replication machine that
catalyzes DNA synthesis.
 In eucaryotes, a special enzyme called telomerase replicates the DNA at the ends of the chromosomes.
 The rare copying mistakes that slip through the DNA replication machinery are dealt with by the mismatch
repair proteins. The overall accuracy of DNA replication, including mismatch repair, is one mistake per 109
nucleotides copied.
 DNA damage caused by unavoidable chemical reactions is repaired by a variety of enzymes that recognize
damaged DNA and excise a short stretch of the DNA strand that contains it. The missing DNA is resynthesized by
a repair DNA polymerase that uses the undamaged strand as a template.
 Nonhomologous end-joining allows the rapid repair of double-strand DNA breaks; the process often alters the
DNA sequence at the site of the repair.
 Homologous recombination can faithfully repair double-strand DNA breaks using a homologous chromosome
sequence as a guide. During meiosis, a related homologous recombination process causes a shuffling of genetic
information that creates DNA molecules with novel sequences.
 Mobile genetic elements, or transposons, move from place to place in the genomes of their hosts, providing a
source of genetic variation.

CHAPTER 7: From DNA to Protein: How Cells Read the Genome

esseNTiAl CONCepTs
 The flow of genetic information in all living cells is DNA Æ RNA Æ protein. The conversion of the genetic
instructions in DNA into RNAs and proteins is termed gene expression.
 To express the genetic information carried in DNA, the nucleotide sequence of a gene is first transcribed into
RNA. Transcription is catalyzed by the enzyme RNA polymerase. Nucleotide sequences in the DNA molecule
indicate to the RNA polymerase where to start and stop transcribing.
 RNA differs in several respects from DNA. It contains the sugar ribose instead of deoxyribose and the base uracil
(U) instead of thymine (T). RNAs in cells are synthesized as single-stranded molecules, which often fold up into
precise three-dimensional shapes.
 Cells make several different functional types of RNAs, including messenger RNA (mRNA), which carries the
instructions for making proteins; ribosomal RNA (rRNA), which is a component of ribosomes; and transfer RNA
(tRNA), which acts as an adaptor molecule in protein synthesis.
 Transcription begins at DNA sites called promoters. To initiate transcription, eucaryotic RNA polymerases
require the assembly of a complex of general transcription factors at the promoter, whereas bacterial RNA
polymerase requires only an additional subunit, called sigma factor.
 In eucaryotic DNA, most genes are composed of a number of smaller coding regions (exons) interspersed with
noncoding regions (introns). When a eucaryotic gene is transcribed from DNA into RNA, both the exons and
introns are copied.
 Introns are removed from the RNA transcripts in the nucleus by the process of RNA splicing. In a reaction
catalyzed by small ribonucleoprotein complexes known as snRNPs, the introns are excised from the RNA and the
exons are joined together.
  Eucaryotic mRNAs go through several additional RNA processing steps before they leave the nucleus, including
RNA capping and polyadenylation. These reactions, along with splicing, take place as the RNA is being
transcribed. The mature mRNA is then transported to the cytoplasm.
 Translation of the nucleotide sequence of mRNA into a protein takes place in the cytoplasm on large
ribonucleoprotein assemblies called ribosomes. As the mRNA is threaded through a ribosome, its message is
translated into protein.
 The nucleotide sequence in mRNA is read in sets of three nucleotides (codons), each codon corresponding to
one amino acid.
 The correspondence between amino acids and codons is specified by the genetic code. The possible
combinations of the 4 different nucleotides in RNA give 64 different codons in the genetic code. Most amino
acids are specified by more than one codon.
 tRNA acts as an adaptor molecule in protein synthesis. Enzymes called aminoacyl-tRNA synthetases link amino
acids to their appropriate tRNAs. Each tRNA contains a sequence of three nucleotides, the anticodon, which
matches a codon in mRNA by complementary base-pairing between codon and anticodon.
 Protein synthesis begins when a ribosome assembles at an initiation codon (AUG) in mRNA, a process that is
regulated by proteins called translation initiation factors. The completed protein chain is released from the
ribosome when a stop codon (UAA, UAG, or UGA) is reached.
 The stepwise linking of amino acids into a polypeptide chain is catalyzed by an rRNA molecule in the large
ribosomal subunit. Thus, the ribosome is an example of a ribozyme, an RNA molecule that can catalyze a
chemical reaction.
 The degradation of proteins in the cell is carefully controlled. Some proteins are degraded in the cytosol by large
protein complexes called proteasomes.
 From our knowledge of present-day organisms and the molecules they contain, it seems likely that living
systems began with the evolution of RNA molecules that could catalyze their own replication. • It has been
proposed that, as cells evolved, the DNA double helix replaced RNA as a more stable molecule for storing
genetic information, and proteins replaced RNAs as major catalytic and structural components. However,
important reactions such as peptide bond formation are still catalyzed by RNA; these are thought to provide a
glimpse into an ancient, RNA-based world.

CHAPTER 8: Control of Gene Expression

ESSENTIAL CONCEPTS
 A typical eucaryotic cell expresses only a fraction of its genes, and the distinct types of cells in multicellular
organisms arise because different sets of genes are expressed as cells differentiate.
 Although all of the steps involved in expressing a gene can in principle be regulated, for most genes the initiation
of transcription is the most important point of control.
 The transcription of individual genes is switched on and off in cells by transcription regulators. These act by
binding to short stretches of DNA called regulatory DNA sequences.
 Although each transcription regulator has unique features, most bind to DNA using one of a small number of
structural motifs. The precise amino acid sequence that is folded into the DNA-binding motif determines the
particular DNA sequence that is recognized.
 In bacteria, transcription regulators usually bind to regulatory DNA sequences close to where RNA polymerase
binds. They can either activate or repress transcription of the gene. In eucaryotes, regulatory DNA sequences are
often separated from the promoter by many thousands of nucleotide pairs.
 Eucaryotic transcription regulators act in two fundamental ways:
(1) they can directly affect the assembly process of RNA polymerase and the general transcription factors at the
promoter, and
(2) they can locally modify the chromatin structure of promoter regions.
 In eucaryotes, the expression of a gene is generally controlled by a combination of transcription regulators.
 In multicellular plants and animals, the production of different transcription regulators in different cell types
ensures the expression of only those genes appropriate to the particular type of cell.
 Cells in multicellular organisms have mechanisms that enable their progeny to ‘remember’ what type of cell they
should be.
  A single transcription regulator, if expressed in the appropriate precursor cell, can trigger the formation of a
specialized cell type or even an entire organ.

CHAPTER 9: How Genes and Genomes Evolve

ESSENTIAL CONCEPTS
 By comparing the nucleotide or amino acid sequences of contemporary organisms we are beginning to
reconstruct how genomes have evolved in the billions of years that have elapsed since the appearance of the
first cells.
 Genetic variation—the raw material for evolutionary change—occurs by a variety of mechanisms that alter the
nucleotide sequences of genomes. These changes range from simple point mutations to larger-scale duplications
and rearrangements.
 Genetic changes that offer an organism a selective advantage or those that are selectively neutral are the most
likely to be perpetuated. Changes that seriously compromise an organism’s fitness are eliminated through
natural selection.
 Gene duplication is one of the most important sources of genetic diversity. Once a gene has been duplicated,
the two copies can accumulate different mutations and thereby diversify to perform different functions. Over
evolutionary time, repeated rounds of gene duplication and divergence can produce large gene families.
 The evolution of new proteins is thought to have been greatly facilitated by the swapping of exons between
genes to create hybrid proteins with new functions.
 The human genome contains 3.2 ¥ 109 nucleotide pairs divided among 22 autosomes and 2 sex chromosomes.
Only a few percent of that DNA codes for proteins and for structural, regulatory, and catalytic RNAs.
 Individual humans differ from one another by an average of 1 nucleotide pair in every 1000; this variation
underlies our individuality and provides the basis for identifying individuals by DNA analysis.
 Comparing genome sequences of different species provides a powerful way to identify genes and to highlight
other functionally important parts of the genome.
 Because related species (such as human and mouse) share many genes in common, evolutionary changes that
affect how these genes are regulated are especially important in understanding the differences between
species.

CHAPTER 10: Analyzing Genes and Genomes

ESSENTIAL CONCEPTS
 Recombinant DNA technology has revolutionized the study of the cell, making it possible for researchers to pick
out any gene at will from the thousands of genes in a cell and to determine the exact molecular structure of the
gene
 A crucial element in this technology is the ability to cut a large DNA molecule into a specific and reproducible set
of DNA fragments by using restriction nucleases, each of which cuts the DNA double helix only at a particular
nucleotide sequence.
 DNA fragments can be separated from one another on the basis of size by gel electrophoresis.
 Nucleic acid hybridization can detect any given DNA or RNA sequence in a mixture of nucleic acid fragments.
This technique relies on the fact that a single strand of DNA or RNA will form a double helix only with another
nucleic acid strand of the complementary nucleotide sequence.
 Single-stranded DNAs of known sequence and labeled with fluorescent dyes or radioisotopes are used as probes
in hybridization reactions.
 Short DNA strands of any sequence can be made by chemical synthesis in the laboratory.
 DNA cloning techniques enable a DNA sequence to be selected from millions of other sequences and produced
in unlimited amounts in pure form.
 DNA fragments can be joined together in vitro by using DNA ligase to form recombinant DNA molecules that are
not found in nature.
 DNA fragments can be maintained and amplified by inserting them into a DNA molecule capable of replication,
such as a plasmid. This recombinant DNA molecule is then introduced into a rapidly dividing host cell, usually a
bacterium, so that the DNA is replicated at each cell division.
 A collection of cloned fragments of chromosomal DNA representing the complete genome of an organism is
known as a genomic library. The library is often maintained as millions of clones of bacteria, each clone carrying
a different DNA fragment.
 cDNA libraries contain cloned DNA copies of the total mRNA of a particular cell type or tissue. Unlike genomic
DNA clones, cloned cDNAs contain predominantly protein-coding sequences; they lack introns, regulatory DNA
sequences, and promoters. They are thus most suitable for use when the cloned gene is to be expressed to
make a protein.
 The polymerase chain reaction (PCR) is a powerful form of DNA amplification that is carried out in vitro using a
purified DNA polymerase. PCR requires prior knowledge of the sequence to be amplified, because two synthetic
oligonucleotide primers must be synthesized that bracket the portion of DNA to be replicated.
 Historically, genes were cloned using hybridization techniques to identify the plasmid carrying the desired
sequence from a DNA library. Today, most genes are cloned using PCR to greatly amplify them and thereby
obtain a specific sequence from a sample of DNA or mRNA.
 Techniques are now available for rapidly determining the nucleotide sequence of any piece of DNA.
 The complete nucleotide sequences of the genomes of hundreds of different organisms have been determined.
These include bacteria, archaea, yeasts, insects, fish, plants, and mammals.
 Bacteria, yeasts, and mammalian cells can be engineered to synthesize large quantities of any protein from any
organism, thus making it possible to study proteins that are otherwise rare or difficult to isolate.
 Using recombinant DNA techniques, a protein can be joined to a molecular tag, such as the green fluorescent
protein (GFP), which allows the tracking of its movement inside the cell. In the case of GFP, the protein can be
monitored over time in living organisms.
 In situ nucleic acid hybridization can be used to detect the precise location of genes in chromosomes, or RNAs in
cells and tissues.
 By presenting a platform for performing a large number of simultaneous hybridization reactions, DNA
microarrays can be used to monitor the expression of tens of thousands of genes at once.
 Cloned genes can be permanently inserted into the genome of a cell or an organism by using recombinant DNA
technology. Cloned DNA can be altered in vitro to create mutant genes that can then be reinserted into a cell or
an organism to study gene function.
 A straightforward strategy for studying the function of a gene is to delete it from the organism’s genome and
then to study the effect of this knockout on the behavior or appearance of the organism.
 The expression of particular genes can be inhibited in cells or organisms by the technique of RNA interference
(RNAi), which prevents an mRNA from being translated into protein.